CN108866152A - A kind of high-throughput quick medicine-sensitive detection kit used for veterinary clinic - Google Patents
A kind of high-throughput quick medicine-sensitive detection kit used for veterinary clinic Download PDFInfo
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Abstract
The invention discloses a kind of high-throughput quick medicine-sensitive detection kits used for veterinary clinic.The kit includes the enriched medium of independent packaging, medicine sensitive detecting plate, reaction solution；The enriched medium is selected from sterile 1640 culture medium or MH culture medium that 1% sterile calf serum and 1 ‰ sterile NAD is added；The medicine sensitive detecting plate is the reaction plate for being coated with several antibiotic, and each hole of reaction plate is coated with a kind of antibiotic, and the peridium concentration of antibiotic is 1-2 times of maximum plasma concentration after antibiotic clinical administration；The resazurin sterile solution that the reaction solution is 0.2%.Drug sensitive test is carried out using kit of the present invention and method, the time of result is short out, and the drug of one-time detection is more, and kit and method are easily operated, does not need specific apparatus and equipment；Test process is liquid phase reactor, more meets physiological environment, practical close to production；Visual result：Blue indicates sensitive, and red indicates insensitive, and for color closer to blue, drug is more sensitive.
Description
Technical field
The invention belongs to detection reagent field for animals, it is related to a kind of using antibiotic drug sensitive detection reagent for veterinary clinic
Box.
Background technique
Antimicrobial play the role of to the control of bacterial infectious disease it is very important, but due to not scientific in breeding process
, the abuse antimicrobial of blindness, many pathogenic bacterias produce drug resistance, so that antimicrobial imitates the control of bacteriosis
Fruit worse and worse, not only causes drug waste, but also delay treatment, causes very huge economic loss to raiser.
In-vitro antibacterial medicament sensitivity test abbreviation drug sensitive test (AST) refers to that measurement drug is antibacterial in vitro or sterilizes
The test of ability.In Vitro Chemosensitivity Test is that veterinary clinic rationally effectively uses antibiotic, understands pathogenic bacteria drug resistance dynamic not
The research technique that can lack.The method being commonly used is that the American National Clinical Laboratory Standard committee (NCCLS) promulgates
Disk diffusion method and agar dilution, although NCCLS method is easily grasped, test process and result judgement be also standardized,
It is that its is complex for operation step, when clinical use takes a long time.In decades always someone drug sensitivity testing in vitro method is explored,
Research, improvement, it would be desirable to find easier, economic, practical method, such as disk diffusion method (K-B method), agar dilution
Method, constant broth dilution method, micro broth dilution method, E measuring method, analytical instrument method and triphenyltetrazolium chloride
(TTC) method etc..But these methods have some disadvantages, for example operating procedure is comparatively laborious, take a long time and can not be primary right
The carry out drug sensitive test etc. of a large amount of antibiotic.In actual production practice, domestic animal morbidity is often relatively anxious, infects comparatively fast, often
Have little time separation of bacterial to identify, be treated above all finding the drug sensitive to pathogenic bacteria with most fast speed, otherwise
Possible drug sensitive experiment is finished not yet, domestic animal large area infection or dead.So develop it is a kind of it is easy to operate, it is time-consuming it is short,
Actual high-throughput susceptibility kit without special instruments and equipment and approach clinic is imperative.Of the invention entirely tested
Journey is liquid phase reactor, more meets physiological environment, practical close to production；The medicine that user can select cost performance high according to result simultaneously
Object is treated and prevented, to reduce the economic loss of user.
Summary of the invention
The purpose of the present invention is being directed to the above-mentioned deficiency of the prior art, a kind of antibiolics used for veterinary clinic is provided
Quick detection kit.
It is a further object of the present invention to provide medicaments insensitive detection methods outside a kind of animal body.
The purpose of the present invention can be achieved through the following technical solutions：
It is a kind of to use antibiotic drug sensitive detection kit, the enriched medium including independent packaging, susceptibility for veterinary clinic
Detection plate, reaction solution；The enriched medium, which is selected from, is added the sterile of 1% sterile calf serum and 1 ‰ sterile NAD
1640 culture mediums；The medicine sensitive detecting plate is the reaction plate for being coated with several antibiotic, and each hole of reaction plate is coated with a kind of anti-
Raw element, the peridium concentration of antibiotic are 1-2 times of maximum plasma concentration after the clinical normal administration of antibiotic；The reaction solution is
0.2% resazurin sterile solution.
The enriched medium is preferably added to 1% sterile calf serum and sterile 1640 culture of 1 ‰ sterile NAD
Base.
Maximum blood medicine on the medicine sensitive detecting plate after the clinical normal administration of the peridium concentration preferred antibiotics of antibiotic
Concentration or twice of maximum plasma concentration.
The further preferred antibiotic clinic intramuscular injection of the peridium concentration of antibiotic or oral administration on the medicine sensitive detecting plate
Maximum plasma concentration or twice of maximum plasma concentration afterwards.
The reaction plate is 96 orifice plates or 48 orifice plates, preferably 96 hole elisa Plates.
The antibiotic include but is not limited to ampicillin sodium, Ceftiofur, Cefquinome sulfate, Cloxacillin Sodium,
Occrycetin, Thiamphenicol, hydrochloric acid sarafloxacin, Danofloxacin mesylate, sulphadiazine, spectinomycin hydrochloride, nefrosulfin
Pyrazine sodium, flumequine, neomycinsulphate, Tilmicosin, fumaric acid tiamulin, Abbott 56619, sulfadimidine,
Florfenicol, Amoxicillin Sodium, Enrofloxacin, Ciprofloxacin, Doxycycline Hyclate, apramycin sulfate, glues Ceftriaxone Sodium
Rhzomorph, amikacin, daimeton, Lincomycin Hydrochloride, scotcil, streptomysin, terramycin, gentamicin, oxygen
It is a variety of in Flucloxacillin, safe ten thousand rhzomorphs.The type of antibiotic can be deleted or be increased according to the actual situation.
With drug normal use dosage (either intramuscular injection still takes orally), on the basis of the Cmax of drug in blood, respectively
It is coated on 96 hole elisa plates with 2 times of Cmax and 1 times of Cmax, is then freeze-dried and medicine sensitive detecting plate is made, signal such as table 1.
1 drug sensitive plate of table is coated with schematic table
The peridium concentration of clinical 35 kinds of common antibiotic, is shown in Table 2.
The peridium concentration of the clinical 34 kinds of common antibiotic of 2 pig of table
A kind of outer medicaments insensitive detection method of animal body uses antibiotic drug sensitive detection examination for veterinary clinic using described
The measurement of agent box progress Susceptibility.
The outer medicaments insensitive detection method of the animal body preferably includes sample inoculation Zengjing Granule after clinic samples
Base, 37 DEG C of culture 2-6h, terminates to cultivate when slightly becoming cloudy with colourless culture medium；Take 100 hole μ L/ of bacterium solution that susceptibility detection is added
The resazurin sterile solution of the reaction solution 0.2% in 50 holes μ L/ is added in plate, 37 DEG C of culture 16h, mixes, and sees after 37 DEG C of culture 1.5h
It examines as a result, blue indicates sensitive, red indicates insensitive, and for color closer to blue, drug is more sensitive.
The outer medicaments insensitive detection method of the animal body, sample are selected from pathological material of disease, excrement or blood；For samples,
Pathological material of disease is acquired using sterile cotton swab, is added in enriched medium；For fecal specimens, 30min is first stood after acquisition, takes supernatant
Liquid 50-100 μ L is added in enriched medium；For blood sample, after standing 30min, takes serum 50-100 μ L to be added and increase bacterium training
It supports in base.
It is of the present invention for veterinary clinic using antibiotic drug sensitive detection kit in vitro detection bacterium to antibiotic
Application in susceptibility.
Beneficial effect：
The present invention selects sterile 1640 culture medium that 1% sterile calf serum and 1 ‰ sterile NAD is added as increasing bacterium
Culture medium can adapt to most of rapid growth of bacteria；Faster compared with LB and TSB culture medium bacterium speed, it is easier to which observation increases
Bacterium reduces color influence caused by result as LB and TSB culture medium.
On the other hand, the present invention is coated with elisa plate with the maximum plasma concentration (Cmax) after drug normal dose use, point
For 1 times of Cmax and 2 times of Cmax (in view of clinically often doubled using antibiotic using), close to veterinary clinic reality.
Drug sensitive test is carried out using kit of the present invention and method, the time of result is short out, interior for 24 hours to obtain result
(except needing bacterium to separate)；The drug of one-time detection is more, and one piece of plank at most can detecte 47 kinds of antibiotic；Kit
It is easily operated, do not need specific apparatus and equipment；Visual result：Blue indicates sensitive, and red indicates insensitive, and color is closer
In blue, drug is more sensitive.
It can also particularly be customized according to the demand of client using the present invention.
Detailed description of the invention
The quick enriched medium of Fig. 1 1640, influence of the LB and TSB culture medium to Actinobacillus enriching effect
Study of Sensitivity of the various antibiotic of Fig. 2 to haemophilus parasuis
Remarks：The medicine name and concentration in each hole, the number in hole are longitudinally A-H, are laterally 1-9 (similarly hereinafter)
A1 ampicillin sodium (14 μ g/mL) B1 ampicillin sodium (7 μ g/mL)
C1 Ceftiofur (24 μ g/mL) D1 Ceftiofur (12 μ g/mL)
E1 Cefquinome sulfate (4 μ g/mL) F1 Cefquinome sulfate (2 μ g/mL)
G1 Cloxacillin Sodium (24 μ g/mL) H1 Cloxacillin Sodium (12 μ g/mL)
A2 occrycetin (14 μ g/mL) B2 occrycetin (7 μ g/mL)
C2 Thiamphenicol (8 μ g/mL) D2 Thiamphenicol (4 μ g/mL)
E2 hydrochloric acid sarafloxacin (2 μ g/mL) F2 hydrochloric acid sarafloxacin (1 μ g/mL)
G2 Danofloxacin mesylate (1 μ g/mL) H2 Danofloxacin mesylate (0.5 μ g/mL)
A3 sulphadiazine (50 μ g/mL) B3 sulphadiazine (25 μ g/mL)
C3 spectinomycin hydrochloride (200 μ g/mL) D3 spectinomycin hydrochloride (100 μ g/mL)
E3 sulfaclozine sodium (100 μ g/mL) F3 sulfaclozine sodium (50 μ g/mL)
G3 flumequine (10 μ g/mL) H3 flumequine (5 μ g/mL)
A4 neomycinsulphate (16 μ g/mL) sulfate framycetin 4 (8 μ g/mL)
C4 Tilmicosin (50 μ g/mL) D4 Tilmicosin (25 μ g/mL)
E4 fumaric acid tiamulin (10 μ g/mL) F4 fumaric acid tiamulin (5 μ g/mL)
G4 Abbott 56619 (4 μ g/mL) H4 Abbott 56619 (2 μ g/mL)
A5 sulfadimidine (100 μ g/mL) B5 sulfadimidine (50 μ g/mL)
C5 Florfenicol (10 μ g/mL) Florfenicol D5 (5 μ g/mL)
E5 ceftriaxone (50 μ g/mL) F5 ceftriaxone (25 μ g/mL)
G5 Amoxicillin Sodium (30 μ g/mL) H5 Amoxicillin Sodium (15 μ g/mL)
A6 Enrofloxacin (1 μ g/mL) B6 Enrofloxacin (0.5 μ g/mL)
C6 Ciprofloxacin (2 μ g/mL) D6 Ciprofloxacin (1 μ g/mL)
E6 Doxycycline Hyclate (4 μ g/mL) F6 Doxycycline Hyclate (2 μ g/mL)
G6 apramycin sulfate (70 μ g/mL) H6 apramycin sulfate (35 μ g/mL)
A7 colistin (4 μ g/mL) B7 colistin (2 μ g/mL)
C7 amikacin (40 μ g/mL) D7 amikacin (20 μ g/mL)
E7 daimeton (30 μ g/mL) F7 daimeton (15 μ g/mL)
G7 Lincomycin Hydrochloride (10 μ g/mL) H7 Lincomycin Hydrochloride (5 μ g/mL)
A8 scotcil (14 μ g/mL) B8 scotcil (7 μ g/mL)
C8 streptomysin (2 μ g/mL) D8 streptomysin (1 μ g/mL)
E8 terramycin (2 μ g/mL) F8 terramycin (1 μ g/mL)
G8 gentamicin (10 μ g/mL) H8 gentamicin (5 μ g/mL)
A9 Ofloxacin (10 μ g/mL) B9 Ofloxacin (5 μ g/mL)
C9 Thailand ten thousand rhzomorphs (4 μ g/mL) D9 ten thousand rhzomorphs of Thailand (2 μ g/mL)
E9 blank F9 blank
Study of Sensitivity of the various antibiotic of Fig. 3 to Actinobacillus
Remarks：The medicine name and concentration in each hole
A1 ampicillin sodium (14 μ g/mL) B1 ampicillin sodium (7 μ g/mL)
C1 Ceftiofur (24 μ g/mL) D1 Ceftiofur (12 μ g/mL)
E1 Cefquinome sulfate (4 μ g/mL) F1 Cefquinome sulfate (2 μ g/mL)
G1 Cloxacillin Sodium (24 μ g/mL) H1 Cloxacillin Sodium (12 μ g/mL)
A2 occrycetin (14 μ g/mL) B2 occrycetin (7 μ g/mL)
C2 Thiamphenicol (8 μ g/mL) D2 Thiamphenicol (4 μ g/mL)
E2 hydrochloric acid sarafloxacin (2 μ g/mL) F2 hydrochloric acid sarafloxacin (1 μ g/mL)
G2 Danofloxacin mesylate (1 μ g/mL) H2 Danofloxacin mesylate (0.5 μ g/mL)
A3 sulphadiazine (50 μ g/mL) B3 sulphadiazine (25 μ g/mL)
C3 spectinomycin hydrochloride (200 μ g/mL) D3 spectinomycin hydrochloride (100 μ g/mL)
E3 sulfaclozine sodium (100 μ g/mL) F3 sulfaclozine sodium (50 μ g/mL)
G3 flumequine (10 μ g/mL) H3 flumequine (5 μ g/mL)
A4 neomycinsulphate (16 μ g/mL) sulfate framycetin 4 (8 μ g/mL)
C4 Tilmicosin (50 μ g/mL) D4 Tilmicosin (25 μ g/mL)
E4 fumaric acid tiamulin (10 μ g/mL) F4 fumaric acid tiamulin (5 μ g/mL)
G4 Abbott 56619 (4 μ g/mL) H4 Abbott 56619 (2 μ g/mL)
A5 sulfadimidine (100 μ g/mL) B5 sulfadimidine (50 μ g/mL)
C5 Florfenicol (10 μ g/mL) Florfenicol D5 (5 μ g/mL)
E5 ceftriaxone (50 μ g/mL) F5 ceftriaxone (25 μ g/mL)
G5 Amoxicillin Sodium (30 μ g/mL) H5 Amoxicillin Sodium (15 μ g/mL)
A6 Enrofloxacin (1 μ g/mL) B6 Enrofloxacin (0.5 μ g/mL)
C6 Ciprofloxacin (2 μ g/mL) D6 Ciprofloxacin (1 μ g/mL)
E6 Doxycycline Hyclate (4 μ g/mL) F6 Doxycycline Hyclate (2 μ g/mL)
G6 apramycin sulfate (70 μ g/mL) H6 apramycin sulfate (35 μ g/mL)
A7 colistin (4 μ g/mL) B7 colistin (2 μ g/mL)
C7 amikacin (40 μ g/mL) D7 amikacin (20 μ g/mL)
E7 daimeton (30 μ g/mL) F7 daimeton (15 μ g/mL)
G7 Lincomycin Hydrochloride (10 μ g/mL) H7 Lincomycin Hydrochloride (5 μ g/mL)
A8 scotcil (14 μ g/mL) B8 scotcil (7 μ g/mL)
C8 streptomysin (2 μ g/mL) D8 streptomysin (1 μ g/mL)
E8 terramycin (2 μ g/mL) F8 terramycin (1 μ g/mL)
G8 gentamicin (10 μ g/mL) H8 gentamicin (5 μ g/mL)
A9 Ofloxacin (10 μ g/mL) B9 Ofloxacin (5 μ g/mL)
C9 Thailand ten thousand rhzomorphs (4 μ g/mL) D9 ten thousand rhzomorphs of Thailand (2 μ g/mL)
E9 blank F9 blank
Study of Sensitivity of the various antibiotic of Fig. 4 to Escherichia coli
Remarks：The medicine name and concentration in each hole
A1 ampicillin sodium (14 μ g/mL) B1 ampicillin sodium (7 μ g/mL)
C1 Ceftiofur (24 μ g/mL) D1 Ceftiofur (12 μ g/mL)
E1 Cefquinome sulfate (4 μ g/mL) F1 Cefquinome sulfate (2 μ g/mL)
G1 Cloxacillin Sodium (24 μ g/mL) H1 Cloxacillin Sodium (12 μ g/mL)
A2 occrycetin (14 μ g/mL) B2 occrycetin (7 μ g/mL)
C2 Thiamphenicol (8 μ g/mL) D2 Thiamphenicol (4 μ g/mL)
E2 hydrochloric acid sarafloxacin (2 μ g/mL) F2 hydrochloric acid sarafloxacin (1 μ g/mL)
G2 Danofloxacin mesylate (1 μ g/mL) H2 Danofloxacin mesylate (0.5 μ g/mL)
A3 sulphadiazine (50 μ g/mL) B3 sulphadiazine (25 μ g/mL)
C3 spectinomycin hydrochloride (200 μ g/mL) D3 spectinomycin hydrochloride (100 μ g/mL)
E3 sulfaclozine sodium (100 μ g/mL) F3 sulfaclozine sodium (50 μ g/mL)
G3 flumequine (10 μ g/mL) H3 flumequine (5 μ g/mL)
A4 neomycinsulphate (16 μ g/mL) sulfate framycetin 4 (8 μ g/mL)
C4 Tilmicosin (50 μ g/mL) D4 Tilmicosin (25 μ g/mL)
E4 fumaric acid tiamulin (10 μ g/mL) F4 fumaric acid tiamulin (5 μ g/mL)
G4 Abbott 56619 (4 μ g/mL) H4 Abbott 56619 (2 μ g/mL)
A5 sulfadimidine (100 μ g/mL) B5 sulfadimidine (50 μ g/mL)
C5 Florfenicol (10 μ g/mL) Florfenicol D5 (5 μ g/mL)
E5 ceftriaxone (50 μ g/mL) F5 ceftriaxone (25 μ g/mL)
G5 Amoxicillin Sodium (30 μ g/mL) H5 Amoxicillin Sodium (15 μ g/mL)
A6 Enrofloxacin (1 μ g/mL) B6 Enrofloxacin (0.5 μ g/mL)
C6 Ciprofloxacin (2 μ g/mL) D6 Ciprofloxacin (1 μ g/mL)
E6 Doxycycline Hyclate (4 μ g/mL) F6 Doxycycline Hyclate (2 μ g/mL)
G6 apramycin sulfate (70 μ g/mL) H6 apramycin sulfate (35 μ g/mL)
A7 colistin (4 μ g/mL) B7 colistin (2 μ g/mL)
C7 amikacin (40 μ g/mL) D7 amikacin (20 μ g/mL)
E7 daimeton (30 μ g/mL) F7 daimeton (15 μ g/mL)
G7 Lincomycin Hydrochloride (10 μ g/mL) H7 Lincomycin Hydrochloride (5 μ g/mL)
A8 scotcil (14 μ g/mL) B8 scotcil (7 μ g/mL)
C8 streptomysin (2 μ g/mL) D8 streptomysin (1 μ g/mL)
E8 terramycin (2 μ g/mL) F8 terramycin (1 μ g/mL)
G8 gentamicin (10 μ g/mL) H8 gentamicin (5 μ g/mL)
A9 Ofloxacin (10 μ g/mL) B9 Ofloxacin (5 μ g/mL)
C9 Thailand ten thousand rhzomorphs (4 μ g/mL) D9 ten thousand rhzomorphs of Thailand (2 μ g/mL)
E9 blank F9 blank
There is No. 0.5 Maxwell turbidity to compare after Fig. 5 APP culture 3h
Remarks：1 is No. 0.5 Maxwell turbidity pipe；2 be APP；3 be blank cultures
Specific embodiment
Embodiment 1
Respectively based on 1640 culture mediums, LB and TSB culture medium after sterilizing, 1% sterile calf serum and 1 ‰ is added
Sterile NAD.The clinical common Escherichia coli of inoculation respectively, haemophilus parasuis (HPS) and Actinobacillus (APP) etc. are research pair
As measuring its OD600 absorbance value every 1h, every test tube is surveyed 3 times, is averaged and draws its growth curve, filter out adaptation
The culture medium of most of rapid growth of bacteria.As shown in Figure 1, quick Zengjing Granule of haemophilus parasuis based on by 1640
Increase bacterium speed faster on base, it is easier to which observation increases bacterium, while reducing the color as LB and TSB culture medium caused by result
It influences.Actinobacillus pleuropneumoniae and haemophilus parasuis trend having the same.It is with 1640 for Escherichia coli
The quick enriched medium on basis is with LB culture medium, between the two without difference.
Embodiment 2
After pig intramuscular administration normal use dosage, on the basis of the Cmax of drug in blood, respectively with 2 times of Cmax and 1
Times Cmax is coated on 96 hole elisa plates, is then freeze-dried and medicine sensitive detecting plate is made.
Using Escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis as object, examined using kit of the present invention
Survey the antibiotics sensitivity of Escherichia coli, actinobacillus pleuropneumoniae and haemophilus parasuis.Escherichia coli, pig pleuropneumonia
Actinobacillus and haemophilus parasuis cultivate 1h on quick enriched medium based on by 1640 respectively, and 3h and 4 hour, so
After take 100 μ L (2 drop) to be added on drug sensitive plate, after 37 DEG C of culture 16h, the 50 μ L (1 of resazurin sterile solution of every hole addition 0.2%
Drop), result is read after 37 DEG C of culture 1.5h.Various antibiotic are to HPS, drug sensitivity tests such as Fig. 2, Fig. 3 of APP and Escherichia coli
With shown in Fig. 4；Wherein for blue to be completely antibacterial, red is not antibacterial.As a result the condition set up is：Negative control is red.Ginseng
According to disk diffusion method, compare the consistency of two methods obtained a result.By comparing quick paper disk method and quick medicine-sensitive plate
Susceptibility results, the match rate of the two are 100%.
Embodiment 3
A, it using sterile cotton swab acquisition pathological material of disease (in order to shorten the incubation time for increasing bacterium, acquisitions more as far as possible), is added and increases bacterium
In culture medium, 37 DEG C of culture 2-6h are carried out down when slightly being become cloudy with colourless culture medium (about No. 0.5 Maxwell turbidity, Fig. 5)
Single stepping；For fecal specimens, 30min is first stood after acquisition, takes supernatant 50-100 μ L (1-2 drop) that enriched medium is added
In；For blood sample, after standing 30min, serum 50-100 μ L (1-2 drop) is taken to be added in enriched medium.
B, cultured 100 μ L of bacterium solution (2 drop), 37 DEG C of culture 16h are added in the every hole for the drug sensitive plate being coated with；
C, drug sensitive plate is taken out, every hole is added 0.2% 50 μ L of resazurin sterile solution (1 drop), reads after 37 DEG C of culture 1.5h
Take result；
D, drug sensitivity tests determine：For blue to be completely antibacterial, red is not antibacterial.As a result the condition set up is：Negative control
For red.
E, result such as Fig. 2-Fig. 4 shows that 24 kinds of drugs are sensitive to selected Escherichia coli, and 6 kinds of drugs are insensitive, 4 kinds of medicines
Object is insensitive under normal use dosage, sensitive after doubling dose；15 kinds of drugs are sensitive to selected APP, and 11 kinds of drugs are not
Sensitivity, 8 kinds of drugs are insensitive under normal use dosage, sensitive after doubling dose；15 kinds of drugs are sensitive to selected HPS,
11 kinds of drugs are insensitive, and 8 kinds of drugs are insensitive under normal use dosage, sensitive after doubling dose.
Claims (10)
1. a kind of use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that the Zengjing Granule including independent packaging
Base, medicine sensitive detecting plate, reaction solution；The enriched medium, which is selected from, is added 1% sterile calf serum and 1 ‰ sterile NAD
Sterile 1640 culture medium or MH culture medium；The medicine sensitive detecting plate is the reaction plate for being coated with several antibiotic, and reaction plate is every
A hole is coated with a kind of antibiotic, and the peridium concentration of antibiotic is 1-2 times of maximum plasma concentration after antibiotic clinical administration；It is described
Reaction solution be 0.2% resazurin sterile solution.
2. according to claim 1 use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that described
Enriched medium is sterile 1640 culture medium that 1% sterile calf serum and 1 ‰ sterile NAD is added.
3. according to claim 1 use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that described
The peridium concentration of antibiotic is that maximum plasma concentration after antibiotic clinical administration or twice of maximum blood medicine are dense on medicine sensitive detecting plate
Degree.
4. according to claim 1 use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that described
The peridium concentration of antibiotic is the maximum plasma concentration after the intramuscular injection of antibiotic clinic or oral administration or twice on medicine sensitive detecting plate
Maximum plasma concentration.
5. according to claim 1 use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that reaction plate
For 96 orifice plates or 48 orifice plates.
6. according to claim 1 use antibiotic drug sensitive detection kit for veterinary clinic, it is characterised in that described
Antibiotic includes ampicillin sodium, Ceftiofur, Cefquinome sulfate, Cloxacillin Sodium, occrycetin, Thiamphenicol, salt
Sour sarafloxacin, Danofloxacin mesylate, sulphadiazine, spectinomycin hydrochloride, sulfaclozine sodium, flumequine, sulfuric acid are newly mould
Element, Tilmicosin, fumaric acid tiamulin, Abbott 56619, sulfadimidine, Florfenicol, Ceftriaxone Sodium, Ah
First between Amdinocillin sodium, Enrofloxacin, Ciprofloxacin, Doxycycline Hyclate, apramycin sulfate, colistin, amikacin, sulfanilamide (SN)
Oxygen pyrimidine, Lincomycin Hydrochloride, scotcil, streptomysin, terramycin, Tiamulin, gentamicin, Ofloxacin, Thailand ten thousand
It is a variety of in rhzomorph.
7. a kind of outer medicaments insensitive detection method of animal body, it is characterised in that use the confession of any of claims 1-6
Veterinary clinic carries out the measurement of Susceptibility using antibiotic drug sensitive detection kit.
8. the outer medicaments insensitive detection method of animal body according to claim 7, it is characterised in that will after being sampled including clinic
Sample inoculation in enriched medium, 37 DEG C culture 2-6 hours, terminate to cultivate when slightly becoming cloudy with colourless culture medium；Take bacterium
Medicine sensitive detecting plate is added in 100 hole μ L/ of liquid, and 37 DEG C are cultivated 16 hours, and the resazurin that the reaction solution 0.2% in 50 holes μ L/ is added is sterile molten
Liquid mixes, and is observed after 37 DEG C of 1.5 h of culture as a result, blue indicates sensitive, red indicates insensitive, and color is closer to indigo plant
Color, drug are more sensitive.
9. the outer medicaments insensitive detection method of animal body according to claim 8, it is characterised in that sample is selected from pathological material of disease, excrement
Or blood；It for samples, is acquired, is added in enriched medium using sterile cotton swab；For fecal specimens, after acquisition first
30min is stood, supernatant 50-100 μ L is taken to be added in enriched medium；Serum is taken after standing 30 min for blood sample
50-100 μ L is added in enriched medium.
10. of any of claims 1-6 detected for veterinary clinic using antibiotic drug sensitive detection kit in vitro
Application in bacterial antibiotic susceptibility.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810576067.8A CN108866152A (en) | 2018-06-06 | 2018-06-06 | A kind of high-throughput quick medicine-sensitive detection kit used for veterinary clinic |
Applications Claiming Priority (1)
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